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Isolating and confirming the Mudrinserted flanking sequences of maize
MuDR exhibits the highest transposition activity and insertional mutagenesis frequency in Mutator (Mu) family. If we isolate the MuDRinsertionspecific flanking sequences (MuDRFs), it will be crucial for using Mu elementmediated mutants. The MuDRTAILPCR system was constructed and optimized using a combination of MuDRTIRnested specific primers and 12 arbitrary degenerate (AD) primers, modified reaction system and procedure and mutant DNA templates of 87 genotypes from M2 or M2:3 families created by crossing the W22::Mu line (active MuDR donor parent) from the UniformMu population with the Zong31 (Z31) line (recipient parent). Here 129 different MuDRFs were acquired by MuDRTAILPCR, accounting for 86.60 % of the total mutantspecific agarose gel bands. In addition, we confirmed the authenticity of the nonredundant flanking sequence amplifications. The amplified nonredundant flanking sequences accounted for 65.12 % of the total MuDRFs, and 88.00 % of the nonredundant MuDRFs were inserted inside the genes. These results show that the MuDRTAILPCR system that we developed can be used for specifically isolating MuDRFs.
Key words: MuDRTAILPCR, MuDR transposon, MuDRinsertion mutant library, Zea mays L.
E-mail: yshtao2015 hotmail.com
|Coded & Designed by Volodymyr Duplij||Modified 08.08.22|